LOCUS       sequence-173216-        6091 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses T4_gp23-mRFP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6091)
  AUTHORS   Myhrvold C, Kotula JW, Hicks WM, Conway NJ, Silver PA
  TITLE     A distributed cell division counter reveals growth dynamics in the 
            gut microbiota.
  JOURNAL   Nat Commun. 2015 Nov 30;6:10039. doi: 10.1038/ncomms10039.
  PUBMED    26615910
REFERENCE   2  (bases 1 to 6091)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2015-11-30"; volume: "6:10039. doi"; pages: " 
            10.1038/ncomms10039"
COMMENT     Sequence Label: pCAM72
FEATURES             Location/Qualifiers
     source          1..6091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..299
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     222..241
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             310..321
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1888..2565
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2583..2654
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2670..2697
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2738..2767
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2783..2800
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2856..2873)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3026..3072
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3114..3131)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3285..3873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3365..3384)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4044..4835,4836..4904))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4667..4686
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4905..5009)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5077..5095
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(5099..5142)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(5201..6079)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSRLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 cccgccattc agagaagaaa ccaattgtcc atattgcatc agacattgcc gtcactgcgt
       61 cttttactgg ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa gcattctgta
      121 acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa agtgtctata atcacggcag
      181 aaaagtccac attgattatt tgcacggcgt cacactttgc tatgccatag catttttatc
      241 cataagatta gcggatctta cctgacgctt tttatcgcaa ctctctactg tttctccata
      301 cccgctagca aagaggagaa aagaatgact atcaaaacta aagctgaact tttgaacaaa
      361 tggaagccat tactggaagg tgaaggttta ccggaaattg ctaatagcaa acaagcgatt
      421 atcgctaaaa tctttgaaaa ccaggaaaaa gatttccaga cagctccgga atataaagac
      481 gaaaaaattg ctaaggcatt cggttctttc ttaacagaag ctgaaatcgg tggtgaccac
      541 ggttacaatg ctaccaacat cgctgcaggt cagacttctg gcgcagtaac tcagattggc
      601 ccagctgtta tgggtatggt acgtcgtgct attcctaacc tgattgcttt cgatatttgt
      661 ggtgttcagc cgatgaacag cccgactggc caggtattcg cactgcgcgc agtatatggt
      721 aaagacccag tggctgccgg tgctaaagaa gcattccacc caatgtatgg tccagatgca
      781 atgttctctg gtcagggtgc tgctaagaaa ttcccagctc tggctgctag cacacaaacc
      841 acagtaggtg atatctatac tcacttcttc caggaaactg gtactgtata tctgcaagct
      901 tctgttcaag taacaatcga tgctggtgcg actgatgctg ctaaattaga tgctgaaatt
      961 aagaaacaaa tggaagctgg tgcactggta gaaatcgctg aaggtatggc tacttctatc
     1021 gctgaactcc aggaaggttt caatggttct accgataacc catggaatga aatgggcttc
     1081 cgtatcgata agcaagttat cgaagctaaa tctcgtcagc tgaaagctgc ttactctatt
     1141 gaattagcac aagacctccg cgctgttcac ggtatggatg ctgatgctga actgtctggt
     1201 attctggcta cagaaattat gctggaaatc aaccgtgaag ttgttgattg gattaactac
     1261 tcagctcagg ttggtaaatc tggtatgacc ctgactccgg gttctaaagc tggtgtattt
     1321 gacttccagg acccaattga tattcgtggt gctcgctggg cgggtgaatc ctttaaagct
     1381 ctgttgttcc agattgacaa agaagcagtt gaaattgctc gtcagaccgg tcgtggtgaa
     1441 ggtaacttca ttatcgcttc ccgtaacgta gttaacgttt tggcttcagt tgataccggc
     1501 atttcttatg ctgcacaggg tctggctacc ggctttagca ctgatactac caagtcagta
     1561 tttgctggtg ttctgggtgg taaataccgc gtatatatcg accagtatgc taaacaggat
     1621 tatttcactg taggttataa aggtccgaac gaaatggatg ctggtattta ctatgctcca
     1681 tatgtagctc tgactccgct gcgtggttcc gatccgaaga acttccaacc ggtaatggga
     1741 ttcaaaactc gttacggtat cggtatcaac ccatttgcag aatccgctgc tcaggctccg
     1801 gcttctcgca tccagagcgg tatgccttct attctgaata gccttggtaa aaacgcttac
     1861 tttagacgtg tatatgttaa aggtatcatg gcgagtagcg aagacgttat caaagagttc
     1921 atgcgtttca aagttcgtat ggaaggttcc gttaacggtc acgagttcga aatcgaaggt
     1981 gaaggtgaag gtcgtccgta cgaaggtacc cagaccgcta aactgaaagt taccaaaggt
     2041 ggtccgctgc cgttcgcttg ggacatcctg tccccgcagt tccagtacgg ttccaaagct
     2101 tacgttaaac acccggctga catcccggac tacctgaaac tgtccttccc ggaaggtttc
     2161 aaatgggaac gtgttatgaa cttcgaagac ggtggtgttg ttaccgttac ccaggactcc
     2221 tccctgcaag acggtgagtt catctacaaa gttaaactgc gtggtaccaa cttcccgtcc
     2281 gacggtccgg ttatgcagaa aaaaaccatg ggttgggaag cttccaccga acgtatgtac
     2341 ccggaagacg gtgctctgaa aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt
     2401 cactacgacg ctgaagttaa aaccacctac atggctaaaa aaccggttca gctgccgggt
     2461 gcttacaaaa ccgacatcaa actggacatc acctcccaca acgaagacta caccatcgtt
     2521 gaacagtacg aacgtgctga aggtcgtcac tccaccggtg cttaataagg atctccaggc
     2581 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     2641 cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt
     2701 ataggatcct aactcgagta atgagaagct tgggcccgaa caaaaactca tctcagaaga
     2761 ggatctgaat agcgccgtcg accatcatca tcatcatcat tgagtttaaa cggtctccag
     2821 cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc
     2881 agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac
     2941 cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat
     3001 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     3061 ctttcgtttt atctgttgtt tgtcggtgaa ctaatttcta gactgcaggc ttcctcgctc
     3121 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3181 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3241 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca caggctccgc
     3301 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3361 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3421 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3481 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3541 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3601 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3661 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3721 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3781 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3841 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3901 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3961 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4021 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4081 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4141 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4201 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4261 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4321 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4381 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4441 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4501 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4561 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4621 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4681 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4741 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4801 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4861 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4921 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4981 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     5041 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcagaat
     5101 ttcagataaa aaaaatcctt agctttcgct aaggatgatt tctggaattc actagtctat
     5161 gctactccat cgagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta
     5221 catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt
     5281 ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg
     5341 cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc
     5401 gccagcttaa gacgctaatc cctaactgct ggcggaaaag acgtgacaga cgcgacggcg
     5461 acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc
     5521 tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa
     5581 tcgcttccat gcgccgcagc aacaattgct caagcagatt tatcgccagc agctccgaat
     5641 agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct
     5701 ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc
     5761 attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct
     5821 ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc
     5881 ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga
     5941 gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct
     6001 caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat
     6061 cattttgcgc ttcagccata cttttcatac t
//
